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First Report of Carrot Cyst Nematode Heterodera carotae in Mexico: Morphological, Molecular Characterization, and Host Range Study

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Fig. 1

A, Carrot field infected with Heterodera carotae; B, Healthy carrot field; C and D, Carrot secondary roots with white females and cysts (indicated by arrows); E and F, Symptoms of an infected carrot. Chlorosis and reddish foliage.
A, Carrot field infected with Heterodera carotae; B, Healthy carrot field; C and D, Carrot secondary roots with white females and cysts (indicated by arrows); E and F, Symptoms of an infected carrot. Chlorosis and reddish foliage.

Fig. 2

Light microscopic photos. A, Cysts; B, Vulval cone. Scale bar: A, 0.5 mm; B, 20 μm.
Light microscopic photos. A, Cysts; B, Vulval cone. Scale bar: A, 0.5 mm; B, 20 μm.

Fig. 3

Scanning electron microscopic photos. A, Entire body of female; B, Vulval cone; C, En face view of male; D, Posterior region of male with bidentate spicules; E, Midbody of J2 with lateral field; F, Tail of J2.
Scanning electron microscopic photos. A, Entire body of female; B, Vulval cone; C, En face view of male; D, Posterior region of male with bidentate spicules; E, Midbody of J2 with lateral field; F, Tail of J2.

Fig. 4

Light microscopic photos. A, Entire body of second stage juvenile (J2); B, Head of J2; C, Tail of J2; D, Head of male; E, Tail with spiclules of male. Scale bar: A, 45 μm; B and C, 25 μm; D and E, 35 μm.
Light microscopic photos. A, Entire body of second stage juvenile (J2); B, Head of J2; C, Tail of J2; D, Head of male; E, Tail with spiclules of male. Scale bar: A, 45 μm; B and C, 25 μm; D and E, 35 μm.

Fig. 5

Phylogenetic relationships within populations and species of the Goettingiana group of the genus Heterodera as inferred from Bayesian analysis using the ITS rRNA gene sequence dataset. Posterior probability more than 70% is given for appropriate clades. Newly obtained sequences are indicated in bold. *Identified as Heterodera goettingiana by Ou et al. (unpublished); **Identified as H. goettingiana by Huang et al. (unpublished); ***Identified as Heterodera sp. by Peng et al. (unpublished) in the GenBank.
Phylogenetic relationships within populations and species of the Goettingiana group of the genus Heterodera as inferred from Bayesian analysis using the ITS rRNA gene sequence dataset. Posterior probability more than 70% is given for appropriate clades. Newly obtained sequences are indicated in bold. *Identified as Heterodera goettingiana by Ou et al. (unpublished); **Identified as H. goettingiana by Huang et al. (unpublished); ***Identified as Heterodera sp. by Peng et al. (unpublished) in the GenBank.

Fig. 6

A, Phylogenetic relationships within populations and species of the Goettingiana group of the genus Heterodera as inferred from Bayesian analysis using the COI gene sequence dataset. Posterior probability more than 70% is given for appropriate clades. B, Statistical parsimony network showing the phylogenetic relationships between haplotypes of species of the Goettingiana group. Small black cycles represent missing haplotypes. Pie chart sizes are proportional to the number of samples with a particular haplotype. Newly obtained sequences are indicated in bold. *Originally identified as Heterodera pratensis by Toumi et al. (2013) and later corrected to Heterodera carotae by Madani et al. (2018).
A, Phylogenetic relationships within populations and species of the Goettingiana group of the genus Heterodera as inferred from Bayesian analysis using the COI gene sequence dataset. Posterior probability more than 70% is given for appropriate clades. B, Statistical parsimony network showing the phylogenetic relationships between haplotypes of species of the Goettingiana group. Small black cycles represent missing haplotypes. Pie chart sizes are proportional to the number of samples with a particular haplotype. Newly obtained sequences are indicated in bold. *Originally identified as Heterodera pratensis by Toumi et al. (2013) and later corrected to Heterodera carotae by Madani et al. (2018).

Morphometrics of J2, males, females, and cysts from Heterodera carotae and H. cruciferae from different populations. All measurements are in micrometer.

Species Heterodera carotae Heterodera cruciferae
Population Characters Mexico (original) Isle of Ely, UK (Jones, 1950b) Ireland, England and Italy (Mathews, 1975) UK, (Stone and Rowe, 1976) Moscow region, Russia (Chizhov et al., 2009) Tabriz, Iran (Jabbari and Niknam, 2008) Spain (Bello et al., 1999)
J2 n = 30 n = 100 n = 25 n = 28 n = 30
Length 394 (360–443) 453.8 ± 1.38 422 (375–452) 431 ± 19 426 ± 31.8 (377–504) 351 ± 15  (333–381) 438 (407–479)
a 18.9 (18.8–20.14) 21.4 (18.2–22.7) 20.7 ± 1.1 20.5 ± 2.7 (14.5–31.8) 20 ± 1.5 (17.5–24) 22.4 (20–25.2)
b 3 (2.99–3.46) 4.5 ± 0.2 (4.1–5.1) 4 ± 0.4 3.4 (2.7–7.4)
b 2.52 (2.5–2.81) 2.4 ± 0.2 (2.1–3.1) 2.4 ± 0.4 (2.06–2.7)
c 8.2 (8.18–8.52) 9.1 ± 0.5 (8.2–10) 8.4 ± 0.7 (7.5–10) 9 (7.9–12.5)
c 3.7 (3.7–4) 3.7 ± 0.3 (3.2–4.4) 4 ± 0.3 (3.5–4.4)
Stylet length 25 (24–26) 24 (22–25) 24.1 ± 1.6 23 ± 0.9 (21–25) 21 ± 0.8 (20–22.4) 23.5 (21.9–25.4)
Labial region height 5 (4–6) 5 (4–6) 4.1 ± 0.7 3–4
Labial region diameter 9 (8–10) 10 (9–10)
DGO 5 (4–6) 5–6 5.5 ± 0.7 3–4 5–6 5.2
Anterior end to valve of median bulb 61 (52–73) 66 (61–72) 68.3 ± 3.1 70 ± 4.1 (62–81)
Anterior end to excretory pore 87 (78–98) 99 101.6 ± 4.8 103 ± 6.2 (91–117) 95–110
Oesophageal length 131 (104–148) 95 ± 5.9 (85–106)
Body diameter at mid-body 21 (19–22) 20 (19–21) 20.8 ± 1.5 21 ± 2.2 (12–27)
Body diameter at anus 13 (11–14) 12.4 (11.1–13.4) 13.6 ± 0.7 13 ± 0.8 (11–14)
Body diameter at hyaline region 8 (7–9) 7.1 (5.9–8.3)
Tail length 48 (44–52) 51.8 (43.5–58.6) 50 ± 2.7 47 ± 4 (38–54) 41 ± 5 (26.2–47) 49.2 (34.1–58.9)
Hyaline region length 26 (21–31) 28.3 (20.3–31.8) 25.2 ± 2.7 24 ± 3.3 (17–30) 21.3 ± 1.9 (16–23.4) 27.3 (20.7–33.9)0.5
Males n = 30 n = 40 n = 25 n = 24 n = 11
Length 1180 (1114–1219) 1119 ± 14 1154 (1090–1220) 1170 ± 79 1156 ± 132.1 (718–1343) 961 ± 83 (828.6–1028.6) 1149 (1081–1283)
a 37 (33.9–42.8) 40.8 ± 5.8 35.3 ± 5.8 (18.9–47.2) 38.2 ± 2.5 (34.1–41.2) 44.5 (34.3 –43.8)
b 6.7 (4.7–7.7) 8.2 ± 0.9 (5.2–9.5) 8.3 ± 0.9 (8.1–10)
b 5.8 (4.6–6.5) 6.1 ± 0.4 (5.2–6.8) 5.3 ± 0.6 (5–6)
Stylet length 32 (29–36) 28.8 ± 0.36 35 (31–38) 24.9 ± 1.1 25 ± 1.7 (22–28) 23 ± 1.6 (20–24.3) 27.1 (25–28)
Labial region height 5 (4–6) 7 6–8
Labial region diameter 11 (9–13) 11 10–12
DGO 5 (4–6) 5–7 7.2 ± 0.8 4–6 6–7
Anterior end to valve of median bulb 103 (101–106) 90 (85–105) 97.8 ± 4.9 96 ± 8.7 (68–107)
Anterior end to excretory pore 157 (146–169) 153 (148–161) 154.4 ± 12.3 159 ± 10.8 (140–182)
Body diameter at mid-body 32 (26–36) 31.4 ± 0.6 30 (27–33) 29.1 ± 1.38 33 ± 5.4 (23–43)
Oesophageal length 175 (144–238) 144 ± 14.3 (119–168)
Anterior end to oesophageal gland base 203 (172–267) 192 ± 14.8 (163–208)
Spicules 36 (32–39) 34 (31–36) 34.5 ± 2.4 34 ± 2.4 (29–38) 21 ± 4.4 (16–26.2) 32.2 (25.4–32.3)
T(%) 48 (39–57) 59 50 48 ± 14.1 (13.5–68)
Females n = 30 n = 10 n = 20
Length 474 (388–686) 451 (375–513) 365.6 ± 28.15
Width 295 (229–430) 203 (168–238) 172.9 ± 18
a 1.6 (1.6–1.7) 2.3 (1.9–2.8)
Stylet length 27 (23–32) 27 (25–31) 26.5 ± 1.5 20.1 ± 4 (15.6–25)
Anterior end to excretory pore 94 (80–116) 81–119 93 ± 20 112.5–143.7
Cysts n = 30 n = 150 n = 25 n = 25 n = 30
Length (excluding neck) 423 (286–575) 210–680 408 (218–625) 429 ± 67 557 ± 71.6 (355–690) 505.4 ± 58.5 (400–609.5) 555 (427–624)
Width 331 (217–472) 150–530 309 (165–500) 333 ± 56 398 ± 72.3 (300–460) 398 ± 63.2(305–571.4) 399 (326–464)
L/W ratio 1.28 (1.22–1.31) 1.4 ± 0.3 (0.6–1.8) 1.3 ± 0.1 (1.1–1.5)
Neck length 80 (63–120) 76 ± 18 66 ± 17.4 (40–90) 66.5 ± 15 (50–84.2)
Fenestral lenth 44 (32–58) 31 (27–36) 34.5 ± 5.9 41 ± 5.5 (27–53) 38.9 (22–58.8)
Fenestral width 50 (38–67) 39 (35–42) 36.6 ± 5 (31.2–44) 38.4 (30–50)
Vulval slit length 48 (39–56) 47 (43–51) 45.5 ± 5.5 46 ± 6.3 (32–55) 42.3 ± 5 (30.2–50) 36.9 (28.8–47.7)

Host range test results of Heterodera carotae from the Tepeaca Valley, Puebla, Mexico.

Latin name Commun name Nematodes
Apiaceae
Daucus carota L. Carrot +
Asteraceae
Lactuca sativa L. Lettuce
Brassicaceae
Brassica oleracea var botrytis L. Cauliflower
Brassica oleracea var capitata L. Cabbage
Brassica oleracea var gemmifera L. Brussels sprouts
Brassica oleracea var italica L. Broccoli
Raphanus sativus L. Radish
Chenopodiaceae
Beta vulgaris L. Beetroot
Poaceae
Triticum aestivum L. Wheat
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